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Transcription factor TFIID recruits factor CPSF for formation of 3′ end of mRNA

Abstract

Initiation of transcription by RNA polymerase II from a promoter region on DNA requires the assembly of several initiation factors to form a preinitiation complex. Assembly of this complex1,2 is initiated by the binding of the transcription factor TFIID, composed of the TATA-box binding protein (TBP) and TBP-associated factors (TAFIIs), to the promoter. We have now characterized an immunopurified TFIID complex which we unexpectedly find contains the cleavage–polyadenylation specificity factor (CPSF), one of the factors required for formation of the 3′ end of messenger RNA3,4. CPSF is brought to the preinitiation complex by TFIID, but after transcription starts, CPSF dissociates from TFIID and becomes associated with the elongating polymerase. We also show that overexpression of recombinant TBP in HeLa cells decreases polyadenylation without affecting the correct initiation of transcription of the reporter gene. This indicates that, owing to incomplete assembly of TFIID on recombinant TBP, CPSF is not brought to the promoter and therefore polyadenylation becomes less efficient. Our observations have thus revealed a link between transcription initiation and elongation by RNA polymerase II and processing of the 3′ end of mRNA.

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Figure 1: CPSF is a component of holo-TFIID.
Figure 2: Interactions of the CPSF-160 with individual components of the TFIID complex.
Figure 3: Fate of CPSF during PIC assembly and transcription.
Figure 4: Overexpressed TBP inhibits polyadenylation of transcripts initiated from the TATA-box-containing promoter of the β-globin gene.
Figure 5: Model depicting the fate of CPSF during transcription initiation, elongation and 3′ pre-mRNA processing.

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References

  1. Roeder, R. G. The role of general initiation factors in transcription by RNA polymerase II. Trends Biochem. Sci. 21, 327–336 (1996).

    Article  CAS  Google Scholar 

  2. Koleske, A. J. & Young, R. A. The RNA polymerase II holoenzyme and its implications for gene regulation. Trends Biochem. Sci. 20, 113–116 (1995).

    Article  CAS  Google Scholar 

  3. Keller, W. No end yet to messenger RNA 3′ processing. Cell 81, 829–832 (1995).

    Article  CAS  Google Scholar 

  4. Manley, J. L. Acomplex protein assembly catalyzes polyadenylation of mRNA precursors. Curr. Opin. Genet. Dev. 5, 222–228 (1995).

    Article  CAS  Google Scholar 

  5. Brou, C. et al. Distinct TFIID complexes mediate the effect of different transcriptional activators. EMBO J. 12, 489–499 (1993).

    Article  CAS  Google Scholar 

  6. Jacq, X. et al. Human TAFII30 is present in a distinct TFIID complex and is required for transcriptional activation by the estrogen receptor. Cell 79, 107–117 (1994).

    Article  CAS  Google Scholar 

  7. Bertolotti, A., Lutz, Y., Heard, D. J., Chambon, P. & Tora, L. hTAFII68, a novel RNA/ssDNA-binding protein with homology to the pro-oncoproteins TLS/FUS and EWS is associated with both TFIID and RNA polymerase II. EMBO J. 15, 5022–5031 (1996).

    Article  CAS  Google Scholar 

  8. Gerard, M. et al. Purification and interaction properties of the human RNA polymerase B(II) general transcription factor BTF2. J. Biol. Chem. 266, 20940–20945 (1991).

    CAS  PubMed  Google Scholar 

  9. Murthy, K. G. & Manley, J. L. The 160-kD subunit of human cleavage-polyadenylation specificity factor coordinates pre-mRNA 3′-end formation. Genes Dev. 9, 2672–2683 (1995).

    Article  CAS  Google Scholar 

  10. Bienroth, S., Wahle, E., Suter-Crazzolara, C. & Keller, W. Purification of the cleavage and polyadenylation factor involved in the 3′-processing of messenger RNA precursors. J. Biol. Chem. 266, 768–776 (1991).

    Google Scholar 

  11. Jenny, A., Minvielle-Sebastia, L., Preker, P. J. & Keller, W. Sequence similarity between the 73-kilodalton protein of mammalian CPSF and a subunit of yeast polyadenylation factor I. Science 274, 1514–1517 (1996).

    Article  ADS  CAS  Google Scholar 

  12. Jenny, A., Hauri, H. P. & Keller, W. Characterization of cleavage and polyadenylation specificity factor and cloning of its 100-kilodalton subunit. Mol. Cell. Biol. 14, 8183–8190 (1994).

    Article  CAS  Google Scholar 

  13. Murthy, K. G. & Manley, J. L. Characterization of the multisubunit cleavage-polyadenylation specificity factor from calf thymus. J. Biol. Chem. 267, 14804–14811 (1992).

    CAS  PubMed  Google Scholar 

  14. Dubrovskaya, V. et al. Distinct domains of hTAFII100 are required for functional interaction with transcription factor TFIIβ (RAP30) and incorporation into the TFIID complex. EMBO J. 15, 3702–3712 (1996).

    Article  CAS  Google Scholar 

  15. Sun, X., Ma, D., Sheldon, M., Yeung, K. & Reinberg, D. Reconstitution of human TFIIA activity from recombinant polypeptides: a role in TFIID-mediated transcription. Genes Dev. 8, 2336–2348 (1994).

    Article  CAS  Google Scholar 

  16. Pugh, B. F. & Tijan, R. Mechanism of transcriptional activation by Sp1: evidence for coactivators. Cell 61, 1187–1197 (1990).

    Article  CAS  Google Scholar 

  17. Zawel, L., Kumar, K. P. & Reinberg, D. Recycling of the general transcription factors during RNA polymerase II transcription. Genes Dev. 9, 1479–1490 (1995).

    Article  CAS  Google Scholar 

  18. Tora, L. et al. The human estrogen receptor has two independent nonacidic transcriptional activation functions. Cell 59, 477–487 (1989).

    Article  CAS  Google Scholar 

  19. Boam, D. S., Davidson, I. & Chambon, P. ATATA-less promoter containing binding sites for ubiquitous transcription factors mediates cell type-specific regulation of the gene for transcription enhancer factor-1 (TEF-1). J. Biol. Chem. 270, 487–494 (1995).

    Article  Google Scholar 

  20. Colgan, J. & Manley, J. L. TFIID can be rate limiting in vivo for TATA-containing, but not TATA-lacking, RNA polymerase II promoters. Genes Dev. 6, 304–315 (1992).

    Article  CAS  Google Scholar 

  21. McCracken, S. et al. The C-terminal domain of RNA polymerase II couples mRNA processing to transcription. Nature 385, 357–361 (1997).

    Article  ADS  CAS  Google Scholar 

  22. Brou, C. et al. Sequence-specific transactivators counteract topoisomerase II-mediated inhibition of in vitro transcription by RNA polymerase I and II. Nucleic Acids Res. 21, 4011–4018 (1993).

    Article  CAS  Google Scholar 

  23. Mengus, G. et al. Cloning and characterization of hTAFII18, hTAFII20 and hTAFII28: three subunits of the human transcription factor TFIID. EMBO J. 14, 1520–1531 (1995).

    Article  CAS  Google Scholar 

  24. May, M., Mengus, G., Lavigne, A.-C., Chambon, P. & Davidson, I. Human TAFII28 promotes transcriptional stimulation by activation function 2 of the retinoid X receptors. EMBO 15, 3093–3104 (1996).

    Article  CAS  Google Scholar 

  25. Lavigne, A. C. et al. Multiple interactions between hTAFII55 and other TFIID subunits. Requirements for the formation of stable ternary complexes between hTAFII55 and the TATA-binding protein. J. Biol. Chem. 271, 774–780 (1996).

    Article  Google Scholar 

  26. Besse, S., Vigneron, M., Pichard, E. & Puvion-Dutilleul, F. Synthesis and maturation of viral transcripts in herpes simplex virus type 1 infected HeLa cells: the role of interchromatin granules. Gene Express. 4, 143–161 (1995).

    CAS  Google Scholar 

  27. Wasylyk, C. & Wasylyk, B. The immunoglobulin heavy-chain B-lymphocyte enhancer efficiently stimulates transcription in non-lymphoid cells. EMBO J. 5, 553–560 (1986).

    Article  CAS  Google Scholar 

  28. Connelly, S. & Manley, J. L. Afunctional mRNA polyadenylation signal is required for transcription termination by RNA polymerase II. Genes Dev. 2, 440–452 (1988).

    Article  CAS  Google Scholar 

  29. Logan, J., Falck-Pedersen, E., Darnell, J. E. J & Shenk, T. Apoly(A) addition site and a downstream termination region are required for efficient cessation of transcription by RNA polymerase II in the mouse beta maj-globin gene. Proc. Natl Acad. Sci. USA 84, 8306–8310 (1987).

    Article  ADS  CAS  Google Scholar 

  30. Whitelaw, E. & Proudfoot, N. Alpha-thalassaemia caused by a poly(A) site mutation reveals that transcriptional termination is linked to 3′ end processing in the human alpha-2 globin gene. EMBO J. 5, 2915–2922 (1986).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank P. Chambon for his continuous support; A. Staub for peptide sequencing; J. M. Chipoulet for DE0.35 fractions; D. Boam, A. Bertolotti, I. Davidson, V. Dubrovskaya, X. Jacq, A.C.Lavigne, G. Mengus and M. Vigneron for reagents; D. J. Heard for critical reading of the manuscript and for discussions; I. Cheynel for expression of the TAFIIs and CPSF in the baculovirus system; Y. Lutz for antibodies; the Cell Culture group for HeLa cells; and R. Buchert and J.-M. Lafontaine for illustrations. J.-C.D. was supported by fellowships from the Ligue National Contre le Cancer and CNRS/Région Alsace. This work was supported by grants from CNRS, INSERM, Centre Hospitalier Universitaire Regional, Ministèe de la Recherche et Technologie, Fondation pour la Recherche Médicale and Association pour la Recherche contre le Cancer (L.T.) and from the NIH (J.L.M.).

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Correspondence to Laszlo Tora.

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Dantonel, JC., Murthy, K., Manley, J. et al. Transcription factor TFIID recruits factor CPSF for formation of 3′ end of mRNA. Nature 389, 399–402 (1997). https://doi.org/10.1038/38763

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